scholarly journals Digital multiplex ligation assay for highly multiplexed screening of β-lactamase-encoding genes in bacterial isolates

2020 ◽  
Vol 3 (1) ◽  
Author(s):  
Manu Tamminen ◽  
Jenny Spaak ◽  
Lea Caduff ◽  
Hanna Schiff ◽  
Ramon Lang ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Target Gene ◽  
Low Cost ◽  
Simultaneous Detection ◽  
Bacterial Isolates ◽  
Cost Efficient ◽  
Encoding Genes ◽  
Multiple Target Gene ◽  
Current Screening ◽  
Digital Multiplex

AbstractIncreasing incidence of antibiotic resistance in clinical and environmental settings calls for increased scalability in their surveillance. Current screening technologies are limited by the number of samples and genes that can easily be screened. We demonstrate here digital multiplex ligation assay (dMLA) as a low-cost targeted genomic detection workflow capable of highly-parallel screening of bacterial isolates for multiple target gene regions simultaneously. Here, dMLA is used for simultaneous detection of 1187 β-lactamase-encoding genes, including extended spectrum β-lactamase (ESBL) genes, in 74 bacterial isolates. We demonstrate dMLA as a light-weight and cost-efficient workflow which provides a highly scalable tool for antimicrobial resistance surveillance and is also adaptable to genetic screening applications beyond antibiotic resistance.

scholarly journals Carbapenemases and extended-spectrum β-lactamases producing Enterobacteriaceae isolated from Tunisian and Libyan hospitals

2016 ◽  
Vol 10 (07) ◽  
pp. 718-727 ◽  
Author(s):  
Najla Mathlouthi ◽  
Charbel Al-Bayssari ◽  
Allaaeddin El Salabi ◽  
Sofiane Bakour ◽  
Salha Ben Gwierif ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Susceptibility Testing ◽  
Test Method ◽  
Bacterial Isolates ◽  
Biochemical Tests ◽  
E Coli ◽  
Extended Spectrum ◽  
Encoding Genes ◽  
Sequence Types ◽  
Esbl Producers

Introduction: The aim of the study was to investigate the prevalence of extended-spectrum β-lactamase (ESBL) and carbapenemase production among clinical isolates of Enterobacteriaceae recovered from Tunisian and Libyan hospitals. Methodology: Bacterial isolates were recovered from patients in intensive care units and identified by biochemical tests and MALDI-TOF. Antibiotic susceptibility testing was performed by disk diffusion and the E-test method. ESBL and carbapenemase activities were detected using standard microbiological tests. Antibiotic resistance-encoding genes were screened by PCR and sequencing. Clonal relationships between Klebsiella pneumoniae strains were carried out using multi-locus sequence typing (MLST). Results: A total of 87 isolates were characterized, with 51 and 36, respectively, identified as E. coli and K. pneumoniae. Overall the resistance prevalence was high for aminoglycosides (> 60%), fluoroquinolones (> 80%), and extended-spectrum cephalosporins (> 94%), and was low for imipenem (11.4%). Among this collection, 58 strains (66.6%) were ESBL producers and 10 K. pneumoniae strains (11.4%) were carbapenemase producers. The antibiotic resistance-encoding genes detected were blaCTX-M-15 (51.7%), blaTEM-1 (35.6%), several variants of blaSHV (21.8%), and blaOXA-48 (11.4%). The MLST typing of K. pneumoniae isolates revealed the presence of multiple clones and three novel sequence types. Also, close relationships between the OXA-48-producing strains from Tunisia and Libya were demonstrated. Conclusions: This study is the first paper describing the emergence of carbapenemase- and ESBL-producing Enterobacteriaceae, sensitive to colistin, isolated in Tunisia and Libya. Active surveillance and testing for susceptibility to colistin should be implementing because resistance to colistin, mainly in Klebsiella, has been recently reported worldwide.

scholarly journals SAM Composition and Electrode Roughness Affect Performance of a DNA Biosensor for Antibiotic Resistance

Biosensors ◽  
2019 ◽  
Vol 9 (1) ◽  
pp. 22 ◽  
Author(s):  
Adrian Butterworth ◽  
Elizabeth Blues ◽  
Paul Williamson ◽  
Milovan Cardona ◽  
Louise Gray ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Target Gene ◽  
Gene Sequence ◽  
Point Of Care ◽  
Low Cost ◽  
Self Assembled Monolayer ◽  
Electrode Surfaces ◽  
Recognition Elements ◽  
Target Gene Sequence ◽  
Electrode Selection

Antibiotic resistance is a growing concern in the treatment of infectious disease worldwide. Point-of-care (PoC) assays which rapidly identify antibiotic resistance in a sample will allow for immediate targeted therapy which improves patient outcomes and helps maintain the effectiveness of current antibiotic stockpiles. Electrochemical assays offer many benefits, but translation from a benchtop measurement system to low-cost portable electrodes can be challenging. Using electrochemical and physical techniques, this study examines how different electrode surfaces and bio-recognition elements, i.e. the self-assembled monolayer (SAM), affect the performance of a biosensor measuring the hybridisation of a probe for antibiotic resistance to a target gene sequence in solution. We evaluate several commercially available electrodes which could be suitable for PoC testing with different SAM layers and show that electrode selection also plays an important role in overall biosensor performance.

The Characterization of Antibiotic Resistance of Bacterial Isolates from Intensive Care Unit Patient Samples in a University Affiliated Hospital in Romania

2019 ◽  
Vol 70 (5) ◽  
pp. 1778-1783
Author(s):  
Andreea-Loredana Golli ◽  
Floarea Mimi Nitu ◽  
Maria Balasoiu ◽  
Marina Alina Lungu ◽  
Cristiana Cerasella Dragomirescu ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Antibiotic Resistance ◽  
Bacterial Pathogens ◽  
Resistance Rate ◽  
Resistance Pattern ◽  
Bacterial Isolates ◽  
E Coli ◽  
Acinetobacter Spp ◽  
Almost All ◽  
Klebsiella Spp

To determine the resistance pattern of bacterial pathogens involved in infections of the patients aged between 18-64 years, admitted in a ICU from a 1518-bed university-affiliated hospital. A retrospective study of bacterial pathogens was carried out on 351 patients aged between 18-64 years admitted to the ICU, from January to December 2017. In this study there were analysed 469 samples from 351 patients (18-64 years). A total of 566 bacterial isolates were obtained, of which 120 strains of Klebsiella spp. (35.39%%), followed by Nonfermenting Gram negative bacilli, other than Pseudomonas and Acinetobacter (NFB) (75- 22.12%), Acinetobacter spp. (53 - 15.63%), Pseudomonas aeruginosa and Proteus (51 - 15.04%), and Escherichia coli (49 - 14.45%). The most common isolates were from respiratory tract (394 isolates � 69.61%). High rates of MDR were found for Pseudomonas aeruginosa (64.70%), MRSA (62.65%) and Klebsiella spp. (53.33%), while almost all of the isolated NFB strains were MDR (97.33%). There was statistic difference between the drug resistance rate of Klebsiella and E. coli strains to ceftazidime and ceftriaxone (p[0.001), cefuroxime (p[0.01) and to cefepime (p[0.01). The study revealed an alarming pattern of antibiotic resistance in the majority of ICU isolates.

scholarly journals Potential impact of biocide adaptation on selection of antibiotic resistance in bacterial isolates

2020 ◽  
Vol 6 (1) ◽  
Author(s):  
Engy Elekhnawy ◽  
Fatma Sonbol ◽  
Ahmed Abdelaziz ◽  
Tarek Elbanna
Keyword(s):  
Antibiotic Resistance ◽  
Cross Resistance ◽  
Main Body ◽  
Bacterial Isolates ◽  
New Agents ◽  
Biocidal Products ◽  
Potential Impact ◽  
Emergence Of Resistance ◽  
Vital Factor ◽  
Selection Of

Abstract Background Antibiotic resistance in pathogenic bacterial isolates has increased worldwide leading to treatment failures. Main body Many concerns are being raised about the usage of biocidal products (including disinfectants, antiseptics, and preservatives) as a vital factor that contributes to the risk of development of antimicrobial resistance which has many environmental and economic impacts. Conclusion Consequently, it is important to recognize the different types of currently used biocides, their mechanisms of action, and their potential impact to develop cross-resistance and co-resistance to various antibiotics. The use of biocides in medical or industrial purposes should be monitored and regulated. In addition, new agents with biocidal activity should be investigated from new sources like phytochemicals in order to decrease the emergence of resistance among bacterial isolates.

scholarly journals Prevalence of Mastitis and Antibiotic Resistance of Bacterial Isolates from CMT Positive Milk Samples Obtained from Dairy Cows, Camels, and Goats in Two Pastoral Districts in Southern Ethiopia

Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1530
Author(s):  
Amanuel Balemi ◽  
Balako Gumi ◽  
Kebede Amenu ◽  
Sisay Girma ◽  
Muuz Gebru ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Dairy Cows ◽  
Raw Milk ◽  
Multidrug Resistant ◽  
Southern Ethiopia ◽  
Resistant Bacteria ◽  
Coagulase Negative Staphylococcus ◽  
Bacterial Isolates ◽  
E Coli ◽  
Milk Samples

A study was carried out from August 2017 to February 2018 on lactating dairy cows, one-humped dromedary camels, and goats to determine mastitis in the Bule Hora and Dugda Dawa districts of in Southern Ethiopia. Milk samples from 564 udder quarters and udder halves from 171 animals consisting of 60 dairy cows, 51 camels, and 60 goats were tested for mastitis. Sixty-four positive udder milk samples were cultured, and bacterial mastitis pathogens were isolated and identified. The antibiotic resistance of bacterial isolates from milk with mastitis was tested against nine antimicrobials commonly used in the study area. Cow- and quarter-level prevalence of mastitis in dairy cows, camels, and goats was 33.3%, 26.3%, and 25% and 17.6%, 14.5%, and 20%, respectively. In cattle, the prevalence was significantly higher in Dugda Dawa than in Bule Hora. Major bacterial isolates were coagulase-negative Staphylococcus species (39.1%), S. aureus (17.2%), S. hyicus (14.1%), and S. intermedius and Escherichia coli (9.4% each). In camels, udder abnormality and mastitis were significantly higher in late lactation than in early lactation. Mastitis tends to increase with parity in camels. E. coli isolates were highly resistant to spectinomycin, vancomycin, and doxycycline, whereas most S. aureus isolates were multidrug-resistant. Most of the rural and periurban communities in this area consume raw milk, which indicates a high risk of infection with multidrug-resistant bacteria. We recommend a community-focused training program to improve community awareness of the need to boil milk and the risk of raw milk consumption.

A simplified low-cost approach to antibiotic resistance profiling for faecal source tracking

2004 ◽  
Vol 50 (1) ◽  
pp. 185-191 ◽  
Author(s):  
J.E. Ebdon ◽  
J.L. Wallis ◽  
H.D. Taylor
Keyword(s):  
Antibiotic Resistance ◽  
Binary Data ◽  
Human Error ◽  
Function Analysis ◽  
Average Rate ◽  
Low Cost ◽  
Source Tracking ◽  
Source Discrimination ◽  
Cost Approach ◽  
Faecal Bacteria

Antibiotic resistance profiling (ARP) is a potentially useful method for distinguishing faecal bacteria according to host source. This phenotypic approach has cost benefits over genotypic methods, but existing protocols are time-consuming and manual data handling is open to human error. A simplified, low-cost approach to the ARP technique was developed that used automated data recording techniques combined with simple statistical analyses to compare isolates of the genus Enterococcus from various faecal sources. An initial battery of 21 antibiotics (at up to four concentrations) was chosen for source discrimination. Images of growth or non-growth in microplate wells were stored as bitmaps and converted to binary data to form a database of known antibiotic resistance profiles. Discriminant function analysis (DFA) showed that the average rate of isolates correctly classified by the database (wastewater vs non-wastewater) was 86%. Once the more discriminating antibiotics and their concentrations had been identified, it was possible to reduce the number of tests from 80 to 18 whilst increasing the number of correctly classified human isolates. ARP could offer a low-cost and rapid means of identifying sources of faecal pollution. As such, the technique may be of particular benefit to developing countries, where water quality may have a significant impact on health and where cost is a major factor when choosing environmental management technology.

Fabrication of hydrophobic NiFe2O4@poly(DVB-LMA) sponge via Pickering emulsion template method for oil/water separation

Soft Matter ◽  
2021 ◽  
Author(s):  
Caimei Zhao ◽  
Lei Chen ◽  
Chuanming Yu ◽  
Binghua Hu ◽  
Haoxuan Huang ◽  
...  
Keyword(s):  
Oil Spills ◽  
Low Cost ◽  
Pickering Emulsion ◽  
Template Method ◽  
Water Separation ◽  
Environment Friendly ◽  
Oil Water Separation ◽  
Oil Water ◽  
Cost Efficient

Super-hydrophobic porous absorbent is a convenient, low-cost, efficient and environment-friendly material in the treatment of oil spills. In this work, a simple Pickering emulsion template method was employed to fabricate...

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